Last data update: 24 January 2024 16:39 CET
Plasmid name: pSEAPEhBCTNf1 (LMBP 3787)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3787.gb
(View with Genome Compiler) p3787.txt p3787.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human placental alkaline phosphatase gene (PLAP, ALPP); mutant encoding a secreted protein (SEAP) |
| Promoter: | Human β-catenin promoter (BCTN, CTNNB1) Simian virus 40 early promoter (SV40 early); enhancer only |
| Ribosome binding site: |
- |
| Terminator: | Simian virus 40 polyadenylation signal (SV40 polyA) |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli Mammalian cells |
| Parental clone: | pSEAPEnhancer; pSEAPBhBCTNf3; pGEM-11Zf(+) |
| Further information: | The plasmid was constructed as follows: 1) the 5,6 kb EcoRI fragment from pSEAPBhBCTNf3 was sheared and resulting fragments were inserted into the unique HindII site of pGEM-11Zf(+); 2) from one of these intermediate constructs an EcoRI/SmaI fragment, containing 103 bp of the human CTNNB1 promoter sequence and 39 bp of the non-coding exon 1 (5' untranslated region) of the human CTNNB1 gene, was isolated and inserted into the Pfu DNA polymerase filled-in HindIII site of pSEAPEnhancer. The SV40 small t-antigen splice signal and the SV40 polyA's downstream from the SEAP gene ensure efficient processing. The SV40 enhancer may increase transcription. The SV40 polyA upstream of the human CTNNB1 promoter reduces background transcription. The nucleotide sequence of this plasmid corresponds with the EMBL Nucleotide Sequence Database accession number LT727392.1. The nucleotide sequence of pSEAPEnhancer corresponds with the EMBL Nucleotide Sequence Database accession number U09662.1. The nucleotide sequence of the human CTNNB1 promoter corresponds with the EMBL Nucleotide Sequence Database accession number X89448.1. Other name of the plasmid is pSEAPEhBCTN-FRAG1. |
| EMBL Accession number: | X89448.1, view at EMBL, GenBank, DDBJ U09662.1, view at EMBL, GenBank, DDBJ LT727392.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 18/07/2008 |
| Sequence detail: | Nucleotide sequence around the termination codon of the human PLAP gene:
485 486 487 488 489 490 491 492 493 513
PLAP: 5' ... GCC GCG CAC CCG GGG CGG TCC GTG GTC......CCC TGA ... 3'
Ala Ala His Pro Gly Arg Ser Val Val Pro *
SEAP: 5' ... GCC GCG CAC CCG GGT TAA CCCGTGGTCCCC ... 3'
Ala Ala His Pro Gly *
HpaI
*: termination codon. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: AvrII/XhoI, BamHI, BglII, SphI and XhoI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr F. Nollet(1) (2) and Prof. Dr F. Van Roy(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Nollet et al., Genomics 32 (1996), 413-424 [PMID: 8838805] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 DH5α |
| Host reference: | Focus 8 (1986), 9 |
| Related host reference: | Woodcock et al., Nucleic Acids Res. 17 (1989), 3469-3478 [PMID: 2657660] Rodriguez-Quinones et al., Focus 15 (1993), 110-112 Hanahan, J. Mol. Biol. 166 (1983), 557-580 [PMID: 6345791] [DOI: 10.1016/s0022-2836(83)80284-8] Hanahan, in 'DNA Cloning: A Practical Approach Volume I', IRL Press, Oxford (1985), 109-135 [ISSN/ISBN: 0947946187] Grant et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 4645-4649 [PMID: 2162051] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pSEAPEhBCTNf1 (LMBP 3787) is available at BCCM/GeneCorner. This plasmid was deposited by Dr F. Nollet and Prof. Dr F. Van Roy . |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.